嗜水气单胞菌外膜蛋白的免疫蛋白质组学研究

Immunoproteomic analysis of outer membrane proteins of Aeromonas hydrophila strain ATCC7966

目的建立嗜水气单胞菌ATCC7966外膜蛋白的免疫蛋白质组学研究方法。方法利用双向电泳技术对嗜水气单胞菌外膜蛋白进行分离，用LC-LTQ-XL质谱技术鉴定，并结合Westernblot技术寻找具有免疫应答的蛋白质。结果LC-LTQ-XL质谱共鉴定出43个肽段，共计39个蛋白质。这些蛋白质不仅有外膜蛋白（69％），还有内膜蛋白（12％）等，功能涉及物质运输、细胞运动和生物合成等领域。最终利用2D结合Westernblot技术找到了一个疫苗候选蛋白。结论成功建立了嗜水气单胞菌外膜蛋白的免疫蛋白质组学研究方法，为将来发现更多候选疫苗提供理论基础。

Objective To identify highly sensitive and specific antigens in Aeromonas hydrophila strain ATCC7966 by using immunoproteomics. Methods Outer membrane proteins were extracted from the Aeromonas hydrophila strain by using two-dimension electrophoresis and identified by LC-LTQ-XL-MS （liq- uid chromatography coupled with linear ion trap mass spectrometry）. Western blot assay was performed to screen out the immunogenic proteins. Results A total of 43 peptides representing 39 proteins were identi- fied by LC-LTQ-XL-MS. Among the 39 proteins, 69% were outer membrane proteins and 12% were inner membrane proteins. They were involved in the process of transportation, cell motility, biosynthesis, etc. One candidate vaccine antigen was identified by using two-dimensional Western blot analysis . Conclusion The immunoproteomics approach could be used to identify immunogenic proteins of Aeromonas hydrophila. This study provided references for further investigation on candidate vaccine antigens.